Abstract
In a recent investigation the author 1 attempted to obtain from spleens of lepers an antigen or product analogous to tuberculin which might be used in the diagnosis of leprosy. The product, obtained by buffer solution extraction of finely ground leprous spleens rich in acid-fast bacilli was tested on leprosy patients by a committee in Manila, P. I. 2 Specific tuberculin-like action could not be demonstrated. There are at least two possible explanations for this negative result. One is that there is no hypersensitiveness to leprosy bacilli. The other is that the acid-fast bacilli in the spleen after the grinding were adsorbed or protected by the preponderantly great amount of normal splenic cells present, and therefore not well extracted.
It seemed preferable, if possible, to separate the bacilli from leprous tissues before grinding and extraction. Previously, attempts have been made to separate acid-fast bacilli from the tissue of the lesions in which they are found. Cowdry 3 and his coworkers dissected bacilli-laden cells from tissues in rat leprosy and then ruptured them with a Mueller press. The expressed tissue fluid, cytoplasm, nucleoplasm and some unruptured cells were ground with sand and centrifuged at low speed. The supernatant fluid, which contained acid-fast bacilli, was then removed and recentrifuged at high speed. The sediment contained a large number of acid-fast organisms. In the experiments here reported advantage was taken of homogenizing chambers 4 for tissue cells and the interface studies of Mudd and Mudd 5 which showed that certain acid-fast organisms, including stock strains of leprosy bacilli, pass readily from the watery into the oil phase.
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